• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.91-100
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• 2003

In the present study, we investigated the protective effect of the Lycii Fructus water extracts (LFE) against CCl4-induced hepatotoxicity and the mechanism underlying these protective effects in the rats. The pretreatment of LFE has shown to possess a significant protective effect by lowering the serum alanine and aspartate aminoteansferase (AST and ALT) and alkaline phosphatase (ALP). This hepatoprotective action was confirmed by histological observation, In addition, the pretreatment of LFE prevented the elevation of hepatic malondialdehyde (MDA) formation and the depletion of reduced glutathione (GSH) content and catalase activity in the liver of CC1₄-injected rats. The LFE also displayed hydroxide radical scavenging activity in a dose-dependent manner (IC50 = 83.6 μg/ml), as assayed by electron spin resonance (ESR) spin-trapping technique. Moreover, the expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR), was significantly decreased in the liver of LFE-pretreated rats when compared with that in the liver of control group. Based on these results, it was suggested that the hepatoprotective effects of the LFE may be related to antioxidant effects and regulation of CYP2E1 gene expression.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.1
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• pp.55-63
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• 1997

The present study was designed to quantify the alterations of renal renin, angiotensin type I receptor ($AT_1$), $TGF-{\beta}1$, and fibronectin gene expression in rats with unilateral ureteral obstruction (UUO). We also investigated the change of $AT_1$ density during UUO. Reverse transcription-polymerase chain reaction (RT-PCR) technique and receptor binding assay were used to detect mRNA expression and receptor density, respectively. At one day after UUO, renin mRNA level of the obstructed kidneys was decreased transiently and then subsequently increased to the level of sham kidneys. In the contralateral kidneys of the same rats, on the contrary, renin mRNA level was gradually decreased. Then, at 9 days after UUO, it was significantly lower than that of sham kidneys. The expressions of both $AT_1$ subtypes, called $AT_{1A}$ and $AT_{1B}$, mRNAs did not change at any time. UUO led to a significant decrease in $AT_1$ density in the obstructed kidneys compared with the sham kidneys at 1 and 3 days $(66\;{\pm}\;11.6%\;(p<0.005)\;and\;73\;{\pm}\;4.0%$ (p<0.01), respectively). Thereafter, $AT_1$ density was gradually increased and at 9 days it showed a marked elevation in the obstructed kidneys compared to the sham kidneys. In contrast, in the contralateral kidneys $AT_1$ density was significantly reduced from 3 to 9 days after UUO. The $TGF-{\beta}$1 mRNA level of the obstructed kidneys was unexpectedly decreased at 6 days after UUO. Then, at 9 days it was followed by a significant increase in the obstructed kidneys, whereas it showed an obvious decrease in the contralateral kidneys. In addition, fibronectin mRNA level was also significantly increased in the obstructed kidneys after UUO compared to the sham or the contralateral kidneys of the same rats. These results suggest a differential regulation of renal renin, $AT_1$ receptor, $TGF-{\beta}$1 and fibronectin mRNA levels at different stages of UUO.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8563-8566
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• 2016

Colorectal cancer is one of the most common lethal cancer types worldwide. In recent years, widespread and large-scale studies have been done on medicinal plants for anti-cancer effects, including Glycyrrhiza glabra. The aim of this study was to evaluate the effects of an ethanol extract Glycyrrhiza glabra on the expression of HSP90, growth and apoptosis in the HT-29 colon cancer cell line. HT-29 cells were treated with different concentrations of extract (50,100,150, and $200{\mu}g/ml$). For evaluation of cell proliferation and apoptosis, we used MTT assay and flow cytometry technique, respectively. RT-PCR was also carried out to evaluate the expression levels of HSP90 genes. Results showed that Glycyrrhiza glabra inhibited proliferation of the HT-29 cell line at a concentration of $200{\mu}g/ml$ and this was confirmed by the highest rate of cell death as measured by trypan blue and MTT assays. RT-PCR results showed down-regulation of HSP90 gene expression which implied an ability of Glycyrrhiza glabra to induce apoptosis in HT-29 cells and confirmed its anticancer property. Further studies are required to evaluate effects of the extract on other genes and also it is necessary to make an extensive in vivo biological evaluation and subsequently proceed with clinical evaluations.

• The Journal of Korean Society of Virology
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• v.28 no.1
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• pp.1-9
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• 1998

To study the relation between ectoparasite, Rickettsia and hanntan virus in bats, four order of Athropoda were collected from Rhinolophus ferrumequinum captured in Cheju and Eptesicus serotinus captured in Chungnamin from July 1989 to March 1998. Also antibody of Rickettsia and hanntan virus were detected by immunofluoroscent antibody technique and RT-PCR. The results are as follows. 1. Five species of Acarina were identified from E. serotinus: Leptotrobidium subakamushi of Trombiculidae, Macronyssus coreanus, Steatonyssuss spinosuss and Steatonychus superans of Macronyssidae, Argas vespertilionis of Metastigmata. 2. Ischnopsyllus needhami of Siphonaptera and Cimex of Hemiptera were identified from E. serotinus. 3. Cycteribia uenoi and Brachytarsina kanoi of Diptera were identified from R. ferrumequinum. 4. The positive rate of rickettsial antibodies in E. serotinus were 17.58%, 15.15%, 22.22%, 52.73% against R. tsutsugamushi, R. typhi, R. sibirica and R. thai tick typhus, respectively. The high positive rate of antibody related to the high content of Arthropoda. 5. The Positive rate of hantaan virus IFA antibodies were 3.4% (27 of 802) and hanntan virus infection rate 36.7% (22 of 60) by RT-PCR in bats. According these result, we showed that certain species of Athropoda isolated playa role as vector of Rickettsia in E. serotinus. Also bats playa role as a reservoir of hantaan virus in nature.

• The Plant Pathology Journal
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• v.33 no.3
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• pp.288-295
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• 2017

Fig mosaic disease (FMD) is a viral disease that spreads in all Tunisian fig (Ficus carica L.) orchards. RT-PCR technique was applied to leaf samples of 29 fig accessions of 15 fig varieties from the fig germplasm collection of High Agronomic Institute (I.S.A) of ChattMariem, to detect viruses associated to FMD. Analysis results show that 65.5% of the accessions (19/29) and 80.0% (12/15) of the fig varieties are infected by FMD-associated viruses. From all fig accessions, 41.4% of them are with single infection (one virus) and 24.1% are with multi-infections (2 virus and more). Viruses infecting fig leaf samples are Fig mosaic virus (FMV) (20.7%), Fig milde-mottle-associated virus (FMMaV) (17.25%), Fig fleck associated virus (FFkaV) (3.45%), and Fig cryptic virus (FCV) (55.17%). A reliable protocol for FCV and FMMaV elimination from 4 local fig varieties Zidi (ZDI), Soltani (SNI), Bither Abiadh (BA), and Assafri (ASF) via in vitro culture of 3 meristem sizes was established and optimized. With this protocol, global sanitation rates of 79.46%, 65.55%, 68.75%, and 70.83% respectively for ZDI, SNI, BA, and ASF are achieved. For all sanitized varieties, the effectiveness of meristem culture for the elimination of FCV and FMMaV viruses was related to meristem size. Meristem size 0.5 mm provides the highest sanitation rates ranging from 70% to 90%.

• Korean Journal of Animal Reproduction
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• v.26 no.4
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• pp.329-338
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• 2002

This study was carried out to investigate the developmental rates of embryo reconstructed with different cell type and to estimate correlation of transcriptional level of octamer-binding transcription factor 4 (Oct4) and fibroblast growth factor 4 (FCF4) gene on peri-implantation stage embryos. Donor cells were transferred into perivitelline space of enucleated oocytes. The karyoplast-cytoplast couplets were accom- plished by cell to cell fusion and activated with ionomycin and 6-dimethylaminopurine. Reconstructed embryos were co-cultured with bovine oviduct epithelial cells in CR 1 aa medium. There is no difference in blastocyst formation rate following nuclear transfer UT) with fetal fibroblast cell (16/50; 32.0%), cumulus cell (16/49; 32.6%) and ear cell (17/52; 32.6%). The expression level of Oct4 and FCF4 in peri-implantation bovine embryo derived from in vitro fertilization (IVF) and NT were determined by reverse-transcription polymerase chain reaction (RT-PCR) technique. In peri-implantation of IVF result in a transient increased of FCF4 paralleled by an increased expression of Oct4. However, Oct4 gene was highly expressed in hatching blastocysts derived from NT compared to IVF. Also, FGF4 expression level in hatching blastocysts and outgrowth stage derived from NT was lower than that of IVF. In conclusion, it is suggested that the different transcription patterns observed in nuclear transfer embryos may lead to a lower rate of embryo development, implantation and pregnancy.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.297-307
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• 2003

In the present study, we investigated the protective effect of the Puerarie Radix water extract (PRE) against CCl₄-induced hepatotoxicity and the mechanism underlying these protective effects in the rats. The pretreatment of PRE has shown to possess a significant protective effect by lowering the serum alanine and aspartate aminoteansferase (AST and ALT) and alkaline phosphatase (ALP). This hepatoprotective action was confirmed by histological observation. In addition, the pretreatment of PRE prevented the elevation of hepatic malondialdehyde (MDA) formation and the depletion of reduced glutathione (GSH) content and catalase activity in the liver of CC1₄-injected rats. The PRE also displayed hydroxide radical scavenging activity in a dose-dependent manner (IC50 = 83.6 μg/ml), as assayed by electron spin resonance (ESR) spin-trapping technique. Moreover, the expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR), was significantly decreased in the liver of PRE-pretreated rats when compared with that in the liver of control group. Based on these results, it was suggested that the hepatoprotective effects of the PRE may be related to antioxidant effects and regulation of CYP2E1 gene expression.

• Korean Journal of Veterinary Service
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• v.27 no.4
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• pp.345-354
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• 2004

Two cases of classical swine fever (CSF) disease have broken out in Cheolwon (7 April, 2002). The suspected pig herds were huddled together because of high fever (over $40^{\circ}C$) and showed remarkable decrease of the leukocytes. The staggering gait related to posterior weakness, constipation and lethargy, hyperemia, hemorrhagic lesions (on the skin, muzzle, ears, limbs, tail and inner part of legs) and conjunctivitis with dirty streaks below the eyes were observed. The inflammation in the lung, infarction in the spleen, swelling and hemorrhage in lymph nodes, kidney, intestine, heart and cheese like purulent inflammation of the tonsil were observed. The ulcers of the colon were also detected. Several clinical and laboratory techniques including blood test, histo-pathological examinations, indirect fluorescent antibody (IFA) test and RT-PCR test were applied to diagnose the disease. Inoculation test on PK-15 cell was also performed. The necrosis of the lymphatic cells and infiltration of the vessel circumferential cells in the brain and lymph organs were commonly viewed. The proliferation of the glia cell (gliosis) in the lymph was particular. Cytopathogenic effect (CPE) and specific fluorescent-bright-green areas (with IFA) appeared in PK-15 cells inoculated with suspected blood plasma. The IFA test on the epithelial and mucous membrane cells of tonsil was positive. RT-PCR technique required more working hours and labor than other techniques in this examination but it was useful because of the sensitivity to the CSF viral gene.

• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.610-620
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• 2021

There has been increasing interest in the head and neck squamous cell carcinoma (HNSCC) that is caused by high-risk human papillomavirus (HR-HPV) and has posed a significant challenge to Otolaryngologists. A rapid, sensitive, and reliable method is required for the detection of HR-HPV in clinical specimens to prevent and treat HPV-induced diseases. In this study, a multiple cross-linking spiral amplification (MCLSA) assay was developed for the visual detection of HPV-16. In the MCLSA assay, samples were incubated under optimized conditions at 62℃ for 45 min, and after mixing with the SYBR Green I (SGI) dye, the positive amplicons showed bright green fluorescence while the negative amplicons exhibited no obvious change. The specificity test revealed that the developed MCLSA technique had high specificity and could effectively distinguish all five HPV-16 strains from other pathogenic microorganisms. In terms of analytical sensitivity, the limit of detection (LoD) of MCLSA assay was approximately 5.4 × 101 copies/tube, which was 10-fold more sensitive than loop-mediated isothermal amplification (LAMP) and RT-PCR. The detection results of laryngeal cancer specimens collected from 46 patients with suspected HPV infection in the Liaoning region demonstrated that the positive detection rates of MCLSA and hybridized capture 2 kit were 32.61% (15/46). The true positive rate of the MCLSA assay was higher than that of RT-PCR (100% vs. 93.33%) and LAMP (100% vs. 86.67%). Therefore, the MCLSA assay developed in the present study could be a potentially useful tool for the point-of-care (PoC) diagnosis of HR-HPV, especially in resource-limited countries.

• BMB Reports
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• v.30 no.4
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• pp.285-291
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• 1997

It is becoming increasingly evident that significant changes in gene expression occur during the course of neuronal differentiation. Thus, it should be possible to gain information about the biochemical events by identifying differentially expressed genes in neuronal differentiation The PC12 cell line is a useful model system to investigate the molecular mechanism underlying neuronal differentiation and has been used extensively for the study of the molecular events that underlie the biological actions of nerve growth factor (NGF). In this study, we report an application of the recently described mRNA differential display method to analyze differential gene expression during neuronal differentiation. Using this technique, we have identified several cDNA tags expressed differentially during neuronal differentiation. Interestingly, one of these clones was cytochrome c oxidase subunit I (COX I) gene. The differential expression of COX I gene was confirmed by Northern blot analysis as well as RT-PCR. Southern blot analysis of the genomic DNA of PC12 cells revealed that COX I is a single gene. Induction of the oxidative enzyme might reflect the energy requirement in neuronal differentiation.